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Isolation and Characterization of the Etheostoma tallapoosae (Teleostei: Percidae) CENP-A Gene.

Fountain DM, Kral LG - Genes (Basel) (2011)

Bottom Line: As a result of this study we have demonstrated that the Tallapoosa darter CENP-A gene HFD sequences can be isolated from genomic DNA by nested PCR in a manner that does not lead to the amplification of the highly sequence related histone H3 gene.These adjacent gene sequences provide us with a primer binding sites for PCR isolation of the CENP-A gene from other percid species of fishes.An initial comparison of three percid species shows that the N-terminal tail of the percid CENP-A gene shows adaptive evolution.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of West Georgia, Carrollton, GA 30118, USA. dfountain82@gmail.com.

ABSTRACT
Both centromeric alpha-satellite sequences as well as centromeric protein A (CENP-A) are highly variable in eukaryotes. CENP-A, a histone H3 variant, is thought to act as the epigenetic "mark" for assembly of centromeric proteins. While most of the histone fold domain (HFD) of the CENP-A is fairly well conserved, a portion of this HFD as well as the N-terminal tail show adaptive variation in both plants and animals. Such variation may establish reproductive barriers that may lead to speciation. The family Percidae contains over 200 species most of which are within the subfamily Etheostomatinae. This subfamily represents a species rich radiation of freshwater fishes in North America and these species exhibit both allopatric and sympatric distributions. In order to study the evolution of CENP-A in percid fish species, we have isolated and characterized the CENP-A gene from Etheostoma tallapoosae by PCR based gene walking. As a result of this study we have demonstrated that the Tallapoosa darter CENP-A gene HFD sequences can be isolated from genomic DNA by nested PCR in a manner that does not lead to the amplification of the highly sequence related histone H3 gene. We also demonstrated that PCR based walking can be subsequently used to isolate the rest of the CENP-A gene and adjacent gene sequences. These adjacent gene sequences provide us with a primer binding sites for PCR isolation of the CENP-A gene from other percid species of fishes. An initial comparison of three percid species shows that the N-terminal tail of the percid CENP-A gene shows adaptive evolution.

No MeSH data available.


Related in: MedlinePlus

Alignment of deduced amino acid sequences of the fish CENP-A protein. Structural and functional features of the HFD are indicated above the alignment according to Regnier et al. [15] and Schueler et al. [14]. Dr: Danio rerio, Ss: Salmo salar, El: Esox lucius, Ga: Gasterosteus aculeatus, Et: Ethesostoma tallapoosae, Fr: Takifugu rubripes, Tn: Tetraodon nigrovirdis.
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f3-genes-02-00829: Alignment of deduced amino acid sequences of the fish CENP-A protein. Structural and functional features of the HFD are indicated above the alignment according to Regnier et al. [15] and Schueler et al. [14]. Dr: Danio rerio, Ss: Salmo salar, El: Esox lucius, Ga: Gasterosteus aculeatus, Et: Ethesostoma tallapoosae, Fr: Takifugu rubripes, Tn: Tetraodon nigrovirdis.

Mentions: In order to carry out a comparison of CENP-A amino acid sequences between the Tallapoosa darter and all other fish species for which the CENP-A sequence has been obtained, we carried out blastp and tblastn searches of GenBank databases with the Tallapoosa darter CENP-A HFD. In addition to the zebrafish and pufferfish sequences previously identified in the databases by Regnier et al. [15], we also identified cDNA sequences from Salmo salar (salmon) and Esox lucius (northern pike). An alignment of all these fish CENP-A sequences is shown in Figure 3. The high degree of homology between the Tallapoosa darter sequence and all the other fish CENP-A sequences confirms that the Tallapoosa darter CENP-A gene has been isolated.


Isolation and Characterization of the Etheostoma tallapoosae (Teleostei: Percidae) CENP-A Gene.

Fountain DM, Kral LG - Genes (Basel) (2011)

Alignment of deduced amino acid sequences of the fish CENP-A protein. Structural and functional features of the HFD are indicated above the alignment according to Regnier et al. [15] and Schueler et al. [14]. Dr: Danio rerio, Ss: Salmo salar, El: Esox lucius, Ga: Gasterosteus aculeatus, Et: Ethesostoma tallapoosae, Fr: Takifugu rubripes, Tn: Tetraodon nigrovirdis.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927593&req=5

f3-genes-02-00829: Alignment of deduced amino acid sequences of the fish CENP-A protein. Structural and functional features of the HFD are indicated above the alignment according to Regnier et al. [15] and Schueler et al. [14]. Dr: Danio rerio, Ss: Salmo salar, El: Esox lucius, Ga: Gasterosteus aculeatus, Et: Ethesostoma tallapoosae, Fr: Takifugu rubripes, Tn: Tetraodon nigrovirdis.
Mentions: In order to carry out a comparison of CENP-A amino acid sequences between the Tallapoosa darter and all other fish species for which the CENP-A sequence has been obtained, we carried out blastp and tblastn searches of GenBank databases with the Tallapoosa darter CENP-A HFD. In addition to the zebrafish and pufferfish sequences previously identified in the databases by Regnier et al. [15], we also identified cDNA sequences from Salmo salar (salmon) and Esox lucius (northern pike). An alignment of all these fish CENP-A sequences is shown in Figure 3. The high degree of homology between the Tallapoosa darter sequence and all the other fish CENP-A sequences confirms that the Tallapoosa darter CENP-A gene has been isolated.

Bottom Line: As a result of this study we have demonstrated that the Tallapoosa darter CENP-A gene HFD sequences can be isolated from genomic DNA by nested PCR in a manner that does not lead to the amplification of the highly sequence related histone H3 gene.These adjacent gene sequences provide us with a primer binding sites for PCR isolation of the CENP-A gene from other percid species of fishes.An initial comparison of three percid species shows that the N-terminal tail of the percid CENP-A gene shows adaptive evolution.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, University of West Georgia, Carrollton, GA 30118, USA. dfountain82@gmail.com.

ABSTRACT
Both centromeric alpha-satellite sequences as well as centromeric protein A (CENP-A) are highly variable in eukaryotes. CENP-A, a histone H3 variant, is thought to act as the epigenetic "mark" for assembly of centromeric proteins. While most of the histone fold domain (HFD) of the CENP-A is fairly well conserved, a portion of this HFD as well as the N-terminal tail show adaptive variation in both plants and animals. Such variation may establish reproductive barriers that may lead to speciation. The family Percidae contains over 200 species most of which are within the subfamily Etheostomatinae. This subfamily represents a species rich radiation of freshwater fishes in North America and these species exhibit both allopatric and sympatric distributions. In order to study the evolution of CENP-A in percid fish species, we have isolated and characterized the CENP-A gene from Etheostoma tallapoosae by PCR based gene walking. As a result of this study we have demonstrated that the Tallapoosa darter CENP-A gene HFD sequences can be isolated from genomic DNA by nested PCR in a manner that does not lead to the amplification of the highly sequence related histone H3 gene. We also demonstrated that PCR based walking can be subsequently used to isolate the rest of the CENP-A gene and adjacent gene sequences. These adjacent gene sequences provide us with a primer binding sites for PCR isolation of the CENP-A gene from other percid species of fishes. An initial comparison of three percid species shows that the N-terminal tail of the percid CENP-A gene shows adaptive evolution.

No MeSH data available.


Related in: MedlinePlus